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Breast Cancer Research and Treatment

, Volume 171, Issue 2, pp 345–357 | Cite as

Syntenin1/MDA-9 (SDCBP) induces immune evasion in triple-negative breast cancer by upregulating PD-L1

  • Jing LiuEmail author
  • Yanfang Yang
  • Hongwei Wang
  • Bin Wang
  • Kaili Zhao
  • Wenna Jiang
  • Weiwei Bai
  • Jun LiuEmail author
  • Jian YinEmail author
Clinical trial

Abstract

Purpose

Syntenin1/SDCBP (syndecan binding protein), also known as melanoma differentiation associated gene-9 (MDA-9), is a PDZ domain-containing molecule, which was initially identified as a key oncogene in melanoma. However, the role of syntenin1 in triple-negative breast cancer (TNBC), especially in suppression of antitumour immune response, remains unknown.

Methods and Results

One hundred TNBC tissues were obtained after radical resection and used for analysis. High syntenin1 expression was associated with increased tumour size (r = 0.421, P < 0.001), presence of lymph node metastasis (r = 0.221, P = 0.044) and poor overall survival (P = 0.01) and recurrence-free survival (P = 0.007). Syntenin1 overexpression significantly promoted 4T1 tumour growth and lung metastasis in BALB/c mice by affecting CD8+ T cells. Western blot and flow cytometry analyses demonstrated that syntenin1 induced CD8+ T cell apoptosis in vitro and in vivo through upregulating PD-L1. Western blot demonstrated that syntenin1 upregulated PD-L1 expression by inducing Tyr705 stat3 phosphorylation, which was further confirmed by stat3 inhibition study. The correlation between syntenin1 and PD-L1 was further confirmed using tumour tissues derived from patients with TNBC (r = 0.509, P < 0.001). Efficacy studies indicated that 4T1-scramble tumour benefitted from anti-PD-L1 therapy (P < 0.001); however, 4T1-syntenin1-KD demonstrated no response to anti-PD-L1 treatment (P = 0.076).

Conclusions

Syntenin1 exhibits a profound function in mediating T cells apoptosis by upregulating PD-L1 and thus could be used as a prognostic biomarker of TNBC. Tumoural syntenin1 expression corelated with anti-PD-L1 treatment efficacy. Targeting syntenin1-mediated T-cell suppression could be a potential strategy for improving the prognosis of patients with TNBC.

Keywords

Triple negative breast cancer Syntenin1 PD-L1 CD8+ T cell apoptosis Immune suppression 

Notes

Funding

The Science &Technology Development Fund of Tianjin Education Commission for Higher Education. (2017KJ198); The Foundation of Tianjin Medical University (2016KYZQ17).

Supplementary material

10549_2018_4833_MOESM1_ESM.docx (14 kb)
Supplementary material 1 (DOCX 13 KB)
10549_2018_4833_MOESM2_ESM.tif (3 mb)
Supplementary Fig1. (a)Syntenin1 protein expression analysis of 4T1-scramble and 4T1-syntenin1-KD by western blotting. 4T1-scramble and 4T1-syntenin1-KD were used for further mouse tumour models. (b) Statistical analysis of the percentage of tumour-infiltrating CD45+ cells. (c) Immunocompromised BABL/c nude mice were inoculated with 4T1-scramble or 4T1-syntenin1-KD in parallel with those immunocompetent WT BABL/c mice. Tumour volumes were measured every 3 days from day 7 to 34 after inoculation. Tumour growth curves for 4T1-scramble groups and 4T1-syntenin1-KD groups were shown. (n=7 per group) * in black P&#x003C;0.05 by repeated measure two-way ANOVA, (time × tumour volume, main effect of group), * in blue P&#x003C;0.05, ** in blue P&#x003C;0.01, by one-way ANOVA followed by Bonferroni post-hoc analysis for each time points.(d) The comparison of tumour volume inhibiting rate after syntenin1 knocked down in WT BABL/c mice and in nude mice. *P&#x003C;0.05 by paired Student’s t-test. (TIF 3115 KB)
10549_2018_4833_MOESM3_ESM.tif (4.3 mb)
Supplementary Fig2. Tumour volumes of 4T1-scramble and 4T1-syntenin1-KD in immunocompetent WT BABL/c mice after receiving CD8+ cells depletion or isotype IgG. (n=10 per group) Results from one-way ANOVA followed by Bonferroni post-hoc analysis for each time points. * P&#x003C;0.05, ** P&#x003C;0.01, ***P&#x003C;0.001 n.s.: not significant. (TIF 4450 KB)
10549_2018_4833_MOESM4_ESM.tif (5.6 mb)
Supplementary Fig3. (a) Statistical analysis of IFNγ secretion. Isolated CD8+T cells were co-cultured with MDA-MB-231-vector, MDA-MB-231-syntenin1, MDA-MB-231-scramble, MDA-MB-231-syntenin1-KD for 48h. ELISA was used to evaluate the concentration of IFNγ in the medium. (b) Statistical analysis of the percentage of activated caspase-3+CD8+T cells. Isolated CD8+T cells were co-cultured with MDA-MB-231-vector, MDA-MB-231-syntenin1, MDA-MB-231-scramble, MDA-MB-231-syntenin1-KD for 12h. Activated caspase-3 and CD8 were labelled and tested by flowcytometry. (c) CFSE dilution of CD8+T cells after 3 days co-cultured with MDA-MB-231-vector, MDA-MB-231-syntenin1, MDA-MB-231-scramble, MDA-MB-231-syntenin1-KD. Proportion of divided CD8+T cells were analysed. *P&#x003C;0.05, **P&#x003C;0.01, by paired Student’s t-test (a, b, c) (TIF 5683 KB)
10549_2018_4833_MOESM5_ESM.tif (4.1 mb)
Supplementary Fig4. Western blot band intensities of syntenin1, PD-L1, P-stat3, t-stat3 relative to β-tubulin were analyzed between groups:(a,c) Statistical analysis of the band intensities ratio between TNBC-vector and TNBC-syntenin1.(b,d) Statistical analysis of the band intensities ratio between TNBC-scramble and TNBC-syntenin1-KD. (TIF 4172 KB)
10549_2018_4833_MOESM6_ESM.tif (4.2 mb)
Supplementary Fig5. Tumour volumes of 4T1-scramble and 4T1-syntenin1-KD in immunocompetent WT BABL/c mice after receiving anti-PD-L1 treatment or isotype IgG. (n=7 per group) Results from one-way ANOVA followed by Bonferroni post-hoc analysis for each time points. ** P&#x003C;0.01, ***P&#x003C;0.001 n.s.: not significant. (TIF 4265 KB)
10549_2018_4833_MOESM7_ESM.tif (1.4 mb)
Supplementary Fig6. Statistical analysis of the percentage of tumour-infiltrating CD45+ cells by one-way ANOVA. (TIF 1473 KB)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Breast Oncoplastic Surgery, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin, Tianjin’s Clinical Research Center for Cancer, Key Laboratory of Breast Cancer Prevention and TherapyTianjin Medical University, Ministry of EducationTianjinChina
  2. 2.Tianjin Medical University, Ministry of EducationTianjinChina
  3. 3.College of Management and EconomicsTianjin UniversityTianjinChina
  4. 4.Department of Laboratory Medicine, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and TherapyTianjin Medical University, Ministry of EducationTianjinChina

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